Spinach hybrid SV2146VB and parents thereof

ABSTRACT

The invention provides seed and plants of spinach hybrid SV2146VB and the parent lines thereof. The invention thus relates to the plants, seeds and tissue cultures of spinach hybrid SV2146VB and the parent lines thereof, and to methods for producing a spinach plant produced by crossing such plants with themselves or with another spinach plant, such as a plant of another genotype. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of such plants, including the leaf and gametes of such plants.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the priority of U.S. Provisional Appl. Ser. No.61/843,787, filed Jul. 8, 2013, the entire disclosure of which isincorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to the field of plant breeding and, morespecifically, to the development of spinach hybrid SV2146VB, and parentspinach lines SSB66-1129F and ESA66-1128M.

BACKGROUND OF THE INVENTION

The goal of vegetable breeding is to combine various desirable traits ina single variety/hybrid. Such desirable traits may include any traitdeemed beneficial by a grower and/or consumer, including greater yield,resistance to insects or pests, tolerance to environmental stress,better agronomic quality, higher nutritional value, growth rate andfruit properties.

Breeding techniques take advantage of a plant's method of pollination.There are two general methods of pollination: a plant self-pollinates ifpollen from one flower is transferred to the same or another flower ofthe same plant or plant variety. A plant cross-pollinates if pollencomes to it from a flower of a different plant variety.

Plants that have been self-pollinated and selected for type over manygenerations become homozygous at almost all gene loci and produce auniform population of true breeding progeny, a homozygous plant. A crossbetween two such homozygous plants of different genotypes produces auniform population of hybrid plants that are heterozygous for many geneloci. Conversely, a cross of two plants each heterozygous at a number ofloci produces a population of hybrid plants that differ genetically andare not uniform. The resulting non-uniformity makes performanceunpredictable.

The development of uniform varieties requires the development ofhomozygous inbred plants, the crossing of these inbred plants, and theevaluation of the crosses. Pedigree breeding and recurrent selection areexamples of breeding methods that have been used to develop inbredplants from breeding populations. Those breeding methods combine thegenetic backgrounds from two or more plants or various other broad-basedsources into breeding pools from which new lines and hybrids derivedtherefrom are developed by selfing and selection of desired phenotypes.The new lines and hybrids are evaluated to determine which of those havecommercial potential.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides a spinach plant of thehybrid designated SV2146VB, the spinach line SSB66-1129F, orESA66-1128M. Also provided are spinach plants having all thephysiological and morphological characteristics of such a plant. Partsof these spinach plants are also provided, for example, includingpollen, an ovule, scion, a rootstock, a fruit, and a cell of the plant.

In another aspect of the invention, a plant of spinach hybrid SV2146VBand/or spinach lines SSB66-1129F and ESA66-1128M comprising an addedheritable trait is provided. The heritable trait may comprise a geneticlocus that is, for example, a dominant or recessive allele. In oneembodiment of the invention, a plant of spinach hybrid SV2146VB and/orspinach lines SSB66-1129F and ESA66-1128M is defined as comprising asingle locus conversion. In specific embodiments of the invention, anadded genetic locus confers one or more traits such as, for example,herbicide tolerance, insect resistance, disease resistance, and modifiedcarbohydrate metabolism. In further embodiments, the trait may beconferred by a naturally occurring gene introduced into the genome of aline by backcrossing, a natural or induced mutation, or a transgeneintroduced through genetic transformation techniques into the plant or aprogenitor of any previous generation thereof. When introduced throughtransformation, a genetic locus may comprise one or more genesintegrated at a single chromosomal location.

The invention also concerns the seed of spinach hybrid SV2146VB and/orspinach lines SSB66-1129F and ESA66-1128M. The spinach seed of theinvention may be provided as an essentially homogeneous population ofspinach seed of spinach hybrid SV2146VB and/or spinach lines SSB66-1129Fand ESA66-1128M. Essentially homogeneous populations of seed aregenerally free from substantial numbers of other seed. Therefore, seedof hybrid SV2146VB and/or spinach lines SSB66-1129F and ESA66-1128M maybe defined as forming at least about 97% of the total seed, including atleast about 98%, 99% or more of the seed. The seed population may beseparately grown to provide an essentially homogeneous population ofspinach plants designated SV2146VB and/or spinach lines SSB66-1129F andESA66-1128M.

In yet another aspect of the invention, a tissue culture of regenerablecells of a spinach plant of hybrid SV2146VB and/or spinach linesSSB66-1129F and ESA66-1128M is provided. The tissue culture willpreferably be capable of regenerating spinach plants capable ofexpressing all of the physiological and morphological characteristics ofthe starting plant, and of regenerating plants having substantially thesame genotype as the starting plant. Examples of some of thephysiological and morphological characteristics of the hybrid SV2146VBand/or spinach lines SSB66-1129F and ESA66-1128M include those traitsset forth in the tables herein. The regenerable cells in such tissuecultures may be derived, for example, from embryos, meristems,cotyledons, pollen, leaves, anthers, roots, root tips, pistils, flowers,seed and stalks. Still further, the present invention provides spinachplants regenerated from a tissue culture of the invention, the plantshaving all the physiological and morphological characteristics of hybridSV2146VB and/or spinach lines SSB66-1129F and ESA66-1128M.

In still yet another aspect of the invention, processes are provided forproducing spinach seeds, plants and fruit, which processes generallycomprise crossing a first parent spinach plant with a second parentspinach plant, wherein at least one of the first or second parentspinach plants is a plant of spinach line SSB66-1129F or ESA66-1128M.These processes may be further exemplified as processes for preparinghybrid spinach seed or plants, wherein a first spinach plant is crossedwith a second spinach plant of a different, distinct genotype to providea hybrid that has, as one of its parents, a plant of spinach lineSSB66-1129F or ESA66-1128M. In these processes, crossing will result inthe production of seed. The seed production occurs regardless of whetherthe seed is collected or not.

In one embodiment of the invention, the first step in “crossing”comprises planting seeds of a first and second parent spinach plant,often in proximity so that pollination will occur for example, mediatedby insect vectors. Alternatively, pollen can be transferred manually.Where the plant is self-pollinated, pollination may occur without theneed for direct human intervention other than plant cultivation.

A second step may comprise cultivating or growing the seeds of first andsecond parent spinach plants into plants that bear flowers. A third stepmay comprise preventing self-pollination of the plants, such as byemasculating the flowers (i.e., killing or removing the pollen).

A fourth step for a hybrid cross may comprise cross-pollination betweenthe first and second parent spinach plants. Yet another step comprisesharvesting the seeds from at least one of the parent spinach plants. Theharvested seed can be grown to produce a spinach plant or hybrid spinachplant.

The present invention also provides the spinach seeds and plantsproduced by a process that comprises crossing a first parent spinachplant with a second parent spinach plant, wherein at least one of thefirst or second parent spinach plants is a plant of spinach hybridSV2146VB and/or spinach lines SSB66-1129F and ESA66-1128M. In oneembodiment of the invention, spinach seed and plants produced by theprocess are first generation (F₁) hybrid spinach seed and plantsproduced by crossing a plant in accordance with the invention withanother, distinct plant. The present invention further contemplatesplant parts of such an F₁ hybrid spinach plant, and methods of usethereof. Therefore, certain exemplary embodiments of the inventionprovide an F₁ hybrid spinach plant and seed thereof.

In still yet another aspect, the present invention provides a method ofproducing a plant derived from hybrid SV2146VB and/or spinach linesSSB66-1129F and ESA66-1128M, the method comprising the steps of: (a)preparing a progeny plant derived from hybrid SV2146VB and/or spinachlines SSB66-1129F and ESA66-1128M, wherein said preparing comprisescrossing a plant of the hybrid SV2146VB and/or spinach lines SSB66-1129Fand ESA66-1128M with a second plant; and (b) crossing the progeny plantwith itself or a second plant to produce a seed of a progeny plant of asubsequent generation. In further embodiments, the method mayadditionally comprise: (c) growing a progeny plant of a subsequentgeneration from said seed of a progeny plant of a subsequent generationand crossing the progeny plant of a subsequent generation with itself ora second plant; and repeating the steps for an additional 3-10generations to produce a plant derived from hybrid SV2146VB and/orspinach lines SSB66-1129F and ESA66-1128M. The plant derived from hybridSV2146VB and/or spinach lines SSB66-1129F and ESA66-1128M may be aninbred line, and the aforementioned repeated crossing steps may bedefined as comprising sufficient inbreeding to produce the inbred line.In the method, it may be desirable to select particular plants resultingfrom step (c) for continued crossing according to steps (b) and (c). Byselecting plants having one or more desirable traits, a plant derivedfrom hybrid SV2146VB and/or spinach lines SSB66-1129F and ESA66-1128M isobtained which possesses some of the desirable traits of the line/hybridas well as potentially other selected traits.

In certain embodiments, the present invention provides a method ofproducing food or feed comprising: (a) obtaining a plant of spinachhybrid SV2146VB and/or spinach lines SSB66-1129F and ESA66-1128M,wherein the plant has been cultivated to maturity, and (b) collecting atleast one spinach from the plant.

In still yet another aspect of the invention, the genetic complement ofspinach hybrid SV2146VB and/or spinach lines SSB66-1129F and ESA66-1128Mis provided. The phrase “genetic complement” is used to refer to theaggregate of nucleotide sequences, the expression of which sequencesdefines the phenotype of, in the present case, a spinach plant, or acell or tissue of that plant. A genetic complement thus represents thegenetic makeup of a cell, tissue or plant, and a hybrid geneticcomplement represents the genetic make up of a hybrid cell, tissue orplant. The invention thus provides spinach plant cells that have agenetic complement in accordance with the spinach plant cells disclosedherein, and seeds and plants containing such cells.

Plant genetic complements may be assessed by genetic marker profiles,and by the expression of phenotypic traits that are characteristic ofthe expression of the genetic complement, e.g., isozyme typing profiles.It is understood that hybrid SV2146VB and/or spinach lines SSB66-1129Fand ESA66-1128M could be identified by any of the many well knowntechniques such as, for example, Simple Sequence Length Polymorphisms(SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990),Randomly Amplified Polymorphic DNAs (RAPDs), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified FragmentLength Polymorphisms (AFLPs) (EP 534 858, specifically incorporatedherein by reference in its entirety), and Single NucleotidePolymorphisms (SNPs) (Wang et al., Science, 280:1077-1082, 1998).

In still yet another aspect, the present invention provides hybridgenetic complements, as represented by spinach plant cells, tissues,plants, and seeds, formed by the combination of a haploid geneticcomplement of a spinach plant of the invention with a haploid geneticcomplement of a second spinach plant, preferably, another, distinctspinach plant. In another aspect, the present invention provides aspinach plant regenerated from a tissue culture that comprises a hybridgenetic complement of this invention.

Any embodiment discussed herein with respect to one aspect of theinvention applies to other aspects of the invention as well, unlessspecifically noted.

The term “about” is used to indicate that a value includes the standarddeviation of the mean for the device or method being employed todetermine the value. The use of the term “or” in the claims is used tomean “and/or” unless explicitly indicated to refer to alternatives onlyor the alternatives are mutually exclusive. When used in conjunctionwith the word “comprising” or other open language in the claims, thewords “a” and “an” denote “one or more,” unless specifically notedotherwise. The terms “comprise,” “have” and “include” are open-endedlinking verbs. Any forms or tenses of one or more of these verbs, suchas “comprises,” “comprising,” “has,” “having,” “includes” and“including,” are also open-ended. For example, any method that“comprises,” “has” or “includes” one or more steps is not limited topossessing only those one or more steps and also covers other unlistedsteps. Similarly, any plant that “comprises,” “has” or “includes” one ormore traits is not limited to possessing only those one or more traitsand covers other unlisted traits.

Other objects, features and advantages of the present invention willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and any specificexamples provided, while indicating specific embodiments of theinvention, are given by way of illustration only, since various changesand modifications within the spirit and scope of the invention willbecome apparent to those skilled in the art from this detaileddescription.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and compositions relating to plants,seeds and derivatives of spinach hybrid SV2146VB, spinach lineSSB66-1129F or ESA66-1128M.

Spinach line ESA66-1128M is also known as ESA-66-1128M.

Spinach hybrid SV2146VB is also known as “RX 066 8 2146”. Spinach(Spinacia oleracea L.) hybrid SV2146VB is a mid early bolting, spinelessseeds (smooth), Savoy leaf-type variety, exhibiting resistance againstDowny mildew Peronospora farinosa f. sp. spinaciae races Pfs 1 till Pfs13. Furthermore the hybrid is phenotypically distinct from allwell-known varieties by a combination of erect petioles, almost roundleafs with a dark, shiny colored leaves.

Spinach (Spinacia oleracea L.) female line SSB 66-1129F exhibits anumber of beneficial traits including resistance to Peronospora farinosaf.sp. spinaciae against the races Pfs 1 to 13 (meanwhile UA0510C isofficially named Pfs13). Line SSB 66-1129F was the subject of the U.S.Ser. No. 13/275,184, and has also been used as a parent to producehybrids as disclosed in U.S. Ser. Nos. 13/781,667, 13/652,248,13/419,260, and 13/436,652.

The development of the line can be summarized as follows.

A. Origin and Breeding History of Spinach Hybrid SV2146VB

Hybrid SV2146VB was developed, in Year 1 at the Monsanto Researchstation in Wageningen, The Netherlands, by crossing female lineSSB66-1129F with male line ESA66-1128M. The final outcome is a mid earlybolting hybrid with Savoy leaves with downy mildew resistances againstthe races Pfs 1 till Pfs 13.

Male line ESAS011-1128M was developed at the Monsanto Research stationin Wageningen, The Netherlands, by pedigree selection out of breedingcross between SVR066 2 2062 and inbred line LSA-66-1044M. The origin andselections that led to the development of ESA66-1128M can be summarizedas follows (S=Selfing, M=Mass selection):

Year Generation Material Main selection criteria Year 1 F1 F1(SVR066 22062* Make cross F1(F1 × Line) LSA-66-1044M) Year 2 F2 Habit (leafshape, color) Year 3 F2.S1 Habit (leaf shape, color) Year 4 F2.S2Habit + Sex expression + Pfs Year 5 F2.S3 Uniformity Year 4 F2.S4Uniformity Year 5 F2.S5 Uniformity Year 6 F2.S6 Uniformity

The final outcome is a line, which combines a mid early bolting habitwith a semi Savoy, oval to pointed leafs and resistance against thedowny mildew races Pfs 1 -7,9,11 and 13. Observation during the Massselection one year and the stock seed production in other yearsconfirmed that ESA66-1128M is uniform and stable. As is true with otherspinach varieties, a small percentage of off-types can occur for almostany characteristics during the course of repeated multiplications.However, no variants were observed during the three years in whichESA66-1128M was observed to be uniform and stable.

Line SSB66-1129F was developed at the Monsanto Research station inWageningen, The Netherlands, by pedigree selection out of a breedingcross in which it was desired to combine a good inbred line with Pfs10resistance from the F1 variety Imola. The origin and selections that ledto the development of SSB66-1129F can be summarized as follows(S=Selfing, M=Mass selection):

Year Generation Material Main selection criteria Year 1 F1[OMB66-1012FxOMB66- Add Pfs10 res. 1018D]*[SVR06894007]* [Imola(PV170)]Year 2 F2. Pfs10 res., color Year 2 F2.S1 Pfs10 res. Year 4 F2.S2.Uniformity Year 5 F2.S2.M1 Uniformity

The final outcome is a line that combines a dark color with theresistance against the races Pfs 1 to Pfs 13. Observation during themass selection one year and the stock seed production in other yearsconfirmed that SMB66-1129F is uniform and stable. As is true with otherspinach varieties, a small percentage of off-types can occur for almostany characteristics during the course of repeated multiplications.However, no variants were observed during the two years in whichSSB66-1129F was observed to be uniform and stable.

Observation during the variety selections in Year 1 until Year 4confirmed that SV2146VB is uniform and stable. As is true with otherspinach varieties, a small percentage of off-types can occur for almostany characteristics during the course of repeated multiplications.However, no variants were observed for SV2146VB. It is uniform andstable. The first test crosses was made in Year 5.

The parent lines are uniform and stable, as is a hybrid producedtherefrom. A small percentage of variants can occur within commerciallyacceptable limits for almost any characteristic during the course ofrepeated multiplication. However no variants are expected.

B. Physiological and Morphological Characteristics of Spinach HybridSv2146vb, and Spinach Lines ssb66-1129f and esa66-1128m

In accordance with one aspect of the present invention, there isprovided a plant having the physiological and morphologicalcharacteristics of spinach hybrid SV2146VB and the parent lines thereof.A description of the physiological and morphological characteristics ofsuch plants is presented in Tables 1-3.

TABLE 1 Physiological and Morphological Characteristics of HybridSV2146VB CHARACTERISTIC SV2146VB Island Ploidy diploid diploid SeedlingCotyledon Length of cotyledon long (Breedblad Scherpzaad, Resistoflay)Width (mm)  6.3 mm  7.2 mm Length (mm) 72.7 mm 50.0 mm Tip pointedpointed Color medium green medium green Color Chart Name RHS Color ChartValue 146A 146B Leaf (First Foliage Leaves) Shape ovate ovate BaseV-shape V-shape Tip round round Margin slightly curled slightly curledUpper Surface Color medium green medium green (Giant Nobel) (GiantNobel) Color Chart Name RHS Color Chart Value 147B 146A Lower SurfaceColor lighter lighter (Compared with upper surface) Color Chart Name RHSColor Chart Value 147C 146B Maturity Growth Rate medium (Long medium(Long Standing Standing Bloomsdale) Bloomsdale) Days from planting to 4343 prime market stage Plant (Prime Market Stage) Habit semi-erect (Longsemi-erect (Long Standing Standing Bloomsdale) Bloomsdale) Size mediummedium Spread (cm) 42.7 cm 41.1 cm Height (cm) 20.0 cm 15.7 cm Leaf(Prime Market Stage) Surface semi-savoy semi-savoy (Northland)(Northland) Blade intensity of green dark [Imola, color Lavewa, Nores]Blade blistering strong [Giraffe, Rhythm] Blade lobing weak [Butterflay,weak [Butterflay, Giraffe] Giraffe] Petiole: attitude semi-erectsemi-erect [Monnopa, Parrot] [Monnopa, Parrot] Petiole: length (primemedium [TG = medium [TG = market stage) Butterflay, Giraffe] Butterflay,Giraffe] Blade attitude horizontal horizontal [Lavewa, Mystic] [Lavewa,Mystic] Blade shape (excluding ovate ovate basal lobes) Blade curving ofmargin flat (Resistoflay) flat (Resistoflay) Blade shape of apex rounded(Imola, rounded (Imola, Nores) Nores) Blade shape in longitudinal convex(Grappa, flat (Mystic, section Lazio) Resistoflay) Base lobed lobed Tipround round Margin flat flat Upper Surface Color dark green (Standingmedium green Bloomsdale) (Giant Nobel) Color Chart Name RHS Color ChartValue 147A 146A Lower Surface Color lighter lighter (Compared with uppersurface) Color Chart Name RHS Color Chart Value 147B 146B Luster dulldull Blade size medium (Virginia medium (Virginia Savoy) Savoy) Petiolecolor medium green medium green Petiole color RHS 143B 143A Petiole redpigmentation absent absent cm petiole length to the 11.2 cm 10.6 cmblade mm petiole diameter  7.5 mm  7.8 mm Petiole diameter medium mediumSeed Stalk Development Start of Bolting (10% of medium (Long medium(Long plants) Standing Standing Bloomsdale) Bloomsdale) Height of Stalk(cm) 83.9 cm 88.1 cm Leaves on Stalk of few or none few or none FemalePlant Leaves on Stalk of Male few or none few or none Plant Plants thatare Female  0-10%  0-10% [Monnopa] [Monnopa] Plants that are Male  0-10% 0-10% [Monnopa, Parrot] [Monnopa, Parrot] Plants that are Monoecious91-100% 91-100% [Monnopa] [Monnopa] Seed Surface smooth smooth Spines(harvested seed) absent [Resistoflay] absent [Resistoflay] *These aretypical values. Values may vary due to environment. Other values thatare substantially equivalent are also within the scope of the invention.

TABLE 2 Physiological and Morphological Characteristics of LineSSB66-1129F CHARACTERISTIC SSB66-1129 F SSB 66-1042F Ploidy diploiddiploid Maturity Growth Rate medium (Long medium (Long StandingBloomsdale) Standing Bloomsdale) Plant (Prime Market Stage) Habit flat(Viroflay) flat (Viroflay) Size medium medium Spread (cm) 40 cm 40 cmHeight (cm) 11 cm 11 cm Seedling Cotyledon Length of cotyledon mediummedium Width (mm)  7 mm  5 mm Length (mm) 62 mm 68 mm Tip roundedrounded Color dark green medium green Color Chart Name RHS Color ChartValue 146A 146B Leaf (First Foliage Leaves) Shape ovate circular BaseV-shape straight Tip round round Margin curled under slightly curledUpper Surface Color dark green (Long dark green (Long StandingBloomsdale) Standing Bloomsdale) Color Chart Name RHS Color Chart Value139A 147A Lower Surface Color lighter lighter (Compared with uppersurface) Color Chart Name RHS Color Chart Value 146A 147B Leaf (PrimeMarket Stage) Surface smooth (Viroflay) smooth (Viroflay) Base lobedlobed Tip round round Margin curled under curled under Upper SurfaceColor dark green (Standing dark green (Standing Bloomsdale) Bloomsdale)Color Chart Name RHS Color Chart Value 147A 147A Lower Surface Colorlighter lighter (Compared with upper surface) Color Chart Name RHS ColorChart Value 146A 146B Luster glossy glossy Blade size medium (Virginiamedium (Virginia Savoy) Savoy) Blade Intensity of green very dark[Lorelay, color mystic] Blade Blistering medium [Butterflay, medium[Butterflay, Koala, Mystic] Koala, Mystic] Blade Lobing absent or veryweak absent or very weak (US = not lobed) (US = not lobed) BladeAttitude horizontal [Lavewa, semi-pendulous Mystic] [Giraffe, Medania]Blade Shape [excluding broad elliptic broad ovate basal lobes] (primemarket stage) Blade curving of margin recurved (Imola) recurved (Imola)Blade shape of apex rounded (Imola, rounded (Imola, Nores) Nores) Bladeshape in (Mystic, Resistoflay) convex (Grappa, longitudinal flat sectionLazio) Petiole Attitude semi-erect [Monnopa, horizontal [Comte, Parrot]Lavewa] Petiole Length (prime medium [TG = long (US = Viroflay) marketstage) Butterflay, Giraffe] [TG = Grappa, Resistoflay] Petiole Colorlight green light green Color Chart Name RHS Color Chart Value 146C 146CPetiole Red Pigmentation absent present (at base) Petiole Length to theBlade  8 cm 10 cm Petiole Diameter (mm)  7 mm  6 mm Petiole Diameterlarge (Giant Nobel) medium Seed Stalk Development Start of Bolting (10%of medium (Long late (Norgreen) plants) Standing Bloomsdale) Time ofstart of bolting late [Grappa, Medania, very late [Chico, (for springsown crop, Revolver] Lavewa] 15% of plants) Height of Stalk (cm) 43 cm51 cm Leaves on Stalk of many many Female Plant Plants that are Female 0-10%  0-10% [Monnopa] [Monnopa] Plants that are Male  0-10%  0-10%[Monnopa, Parrot] [Monnopa, Parrot] Plants that are 91-100% 91-100%Monoecious [Monnopa] [Monnopa] Seed Surface smooth smooth Spines(harvested seed) absent [Resistoflay] absent [Resistoflay] *These aretypical values. Values may vary due to environment. Other values thatare substantially equivalent are also within the scope of the invention.

TABLE 3 Physiological and Morphological Characteristics of LineESA-66-1128M CHARACTERISTIC ESA-66-1128M ESA-66-1070M Ploidy diploiddiploid Maturity Growth Rate medium (Long medium (Long StandingBloomsdale) Standing Bloomsdale) Number days from 40 37 planting toprime market stage Plant (Prime Market Stage) Habit erect (VirginiaSavoy) erect (Virginia Savoy) Size small (America) small (America)Spread (cm) 29.8 cm 24.2 cm Height (cm) 14.4 cm 14.6 cm SeedlingCotyledon Length of cotyledon medium medium Width (mm)  5.8 mm  5.4 mmLength (mm) 53.4 mm 60.5 mm Tip pointed pointed Color dark green darkgreen Color Chart Name RHS Color Chart Value 146A 146A Leaf (FirstFoliage Leaves) Shape ovate circular; ovate Base V-shape straight Tipround round Margin slightly curled slightly curled/curled under UpperSurface Color dark green (Long dark green (Long Standing Bloomsdale)Standing Bloomsdale) Color Chart Name RHS Color Chart Value 147A 146ALower Surface Color lighter lighter (Compared with upper surface) ColorChart Name RHS Color Chart Value 147B 146B Leaf (Prime Market Stage)Surface savoy (Virginia savoy (Virginia Savoy) Savoy) Base lobed lobedTip round round Margin flat flat Upper Surface Color dark green(Standing dark green (Standing Bloomsdale) Bloomsdale) Color Chart NameRHS Color Chart Value 147A 147A Lower Surface Color lighter lighter(Compared with upper surface) Color Chart Name RHS Color Chart Value146A 146A Luster glossy glossy Blade size medium (Virginia medium(Virginia Savoy) Savoy) Blade Intensity of green very dark (Lorelay,dark (Imola, Lavewa, color Mystic) Nores) Blade Lobing weak [Butterflay,absent or very weak Giraffe] (US = not lobed) Blade Attitude horizontal[Lavewa, horizontal [Lavewa, Mystic] Mystic] Blade Shape [excludingovate ovate basal lobes] (prime market stage) Blade curving of marginflat (Resistoflay) flat (Resistoflay) Blade shape of apex rounded(Imola, rounded (Imola, Nores) Nores) Blade shape in convex (Grappa,convex (Grappa, longitudinal section Lazio) Lazio) Petiole Attitudeerect [Grappa] erect [Grappa] Petiole Length (prime medium [TG = medium[TG = market stage) Butterflay, Giraffe] Butterflay, Giraffe]; short [TG= Imola, Mystic] Petiole Color medium green medium green Color ChartName RHS Color Chart Value 143A 143A Petiole Red Pigmentation absentabsent Petiole Length to the Blade  9.4 cm  8.7 cm Petiole Diameter (mm) 6.7 mm  6.9 mm Petiole Diameter medium medium Seed Stalk DevelopmentStart of Bolting (10% of medium (Long medium (Long plants) StandingBloomsdale) Standing Bloomsdale) Time of start of bolting medium[Matador, medium [Matador, (for spring sown crop, Monnopa] Monnopa] 15%of plants) Height of Stalk (cm) 91.1 cm 75.7 cm Plants that are Female 0-10%  0-10% [Monnopa] [Monnopa] Plants that are Male  0-10%  0-10%[Monnopa, Parrot] [Monnopa, Parrot] Plants that are 91-100% 91-100%Monoecious [Monnopa] [Monnopa] Seed Surface smooth smooth Spines(harvested seed) absent [Resistoflay] absent [Resistoflay] *These aretypical values. Values may vary due to environment. Other values thatare substantially equivalent are also within the scope of the invention.

C. Breeding Spinach Plants

One aspect of the current invention concerns methods for producing seedof spinach hybrid SV2146VB involving crossing spinach lines SSB66-1129Fand ESA66-1128M. Alternatively, in other embodiments of the invention,hybrid SV2146VB, line SSB66-1129F or line ESA66-1128M may be crossedwith itself or with any second plant. Such methods can be used forpropagation of hybrid SV2146VB and/or the spinach lines SSB66-1129F andESA66-1128M, or can be used to produce plants that are derived fromhybrid SV2146VB and/or the spinach lines SSB66-1129F and ESA66-1128M.Plants derived from hybrid SV2146VB and/or the spinach lines SSB66-1129Fand ESA66-1128M may be used, in certain embodiments, for the developmentof new spinach varieties.

The development of new varieties using one or more starting varieties iswell known in the art. In accordance with the invention, novel varietiesmay be created by crossing hybrid SV2146VB followed by multiplegenerations of breeding according to such well known methods. Newvarieties may be created by crossing with any second plant. In selectingsuch a second plant to cross for the purpose of developing novel lines,it may be desired to choose those plants which either themselves exhibitone or more selected desirable characteristics or which exhibit thedesired characteristic(s) when in hybrid combination. Once initialcrosses have been made, inbreeding and selection take place to producenew varieties. For development of a uniform line, often five or moregenerations of selfing and selection are involved.

Uniform lines of new varieties may also be developed by way ofdouble-haploids. This technique allows the creation of true breedinglines without the need for multiple generations of selfing andselection. In this manner true breeding lines can be produced in aslittle as one generation. Haploid embryos may be produced frommicrospores, pollen, anther cultures, or ovary cultures. The haploidembryos may then be doubled autonomously, or by chemical treatments(e.g. colchicine treatment). Alternatively, haploid embryos may be growninto haploid plants and treated to induce chromosome doubling. In eithercase, fertile homozygous plants are obtained. In accordance with theinvention, any of such techniques may be used in connection with a plantof the invention and progeny thereof to achieve a homozygous line.

Backcrossing can also be used to improve an inbred plant. Backcrossingtransfers a specific desirable trait from one inbred or non-inbredsource to an inbred that lacks that trait. This can be accomplished, forexample, by first crossing a superior inbred (A) (recurrent parent) to adonor inbred (non-recurrent parent), which carries the appropriate locusor loci for the trait in question. The progeny of this cross are thenmated back to the superior recurrent parent (A) followed by selection inthe resultant progeny for the desired trait to be transferred from thenon-recurrent parent. After five or more backcross generations withselection for the desired trait, the progeny have the characteristicbeing transferred, but are like the superior parent for most or almostall other loci. The last backcross generation would be selfed to givepure breeding progeny for the trait being transferred.

The plants of the present invention are particularly well suited for thedevelopment of new lines based on the elite nature of the geneticbackground of the plants. In selecting a second plant to cross withSV2146VB and/or spinach lines SSB66-1129F and ESA66-1128M for thepurpose of developing novel spinach lines, it will typically bepreferred to choose those plants which either themselves exhibit one ormore selected desirable characteristics or which exhibit the desiredcharacteristic(s) when in hybrid combination. Examples of desirabletraits may include, in specific embodiments, high seed yield, high seedgermination, seedling vigor, high fruit yield, disease tolerance orresistance, and adaptability for soil and climate conditions.Consumer-driven traits, such as a fruit shape, color, texture, and tasteare other examples of traits that may be incorporated into new lines ofspinach plants developed by this invention.

D. Performance Characteristics

As described above, hybrid SV2146VB exhibits desirable traits, asconferred by spinach lines SSB66-1129F and ESA66-1128M. The performancecharacteristics of hybrid SV2146VB and spinach lines SSB66-1129F andESA66-1128M were the subject of an objective analysis of the performancetraits relative to other varieties. The results of the analysis arepresented below.

Spinach line SSB66-1129F can be characterized as a late bolting linethat has spineless seeds (smooth), with dark color and with a resistanceto downy mildew (Peronospora farinose f. sp. spinaciae (Pfs)) races Pfs1 till Pfs 12 and to UA0510C. Furthermore the line is phenotypicallydistinct from all well-known material by a combination slow growing,erect growth habit and dark green leaf color.

The parental line believed to most closely resemble SSB66-1129F isMonsanto parental line SMB66-1100M. Comparative characteristicsdistinguish the two lines that include, but may not be limited tobolting and flowering. The candidate line SSB66-1129F is later boltingand is more female flowering compared to SMB66-1100M.

TABLE 4 Comparison of Hybrid SV2146VB with ISLAND. Location: Wageningen,the Netherlands. Resistances Leaf Color SV2146VB Pfs 1-13 Darker ISLANDPfs 1-11, 13 Lighter

TABLE 5 Performance Characteristics For Line SSB 66-1129F andComparative Variety SMB66-1100M Bolting * Material 2009 2010 2011Flowering Leaf Color SSB66-1129F 31 44 31 Female Darker SMB66-1100M 2532 21 Male Lighter * bolting is number of days after 1^(st) of May

E. Further Embodiments of the Invention

In certain aspects of the invention, plants described herein areprovided modified to include at least a first desired heritable trait.Such plants may, in one embodiment, be developed by a plant breedingtechnique called backcrossing, wherein essentially all of themorphological and physiological characteristics of a variety arerecovered in addition to a genetic locus transferred into the plant viathe backcrossing technique. The term single locus converted plant asused herein refers to those spinach plants which are developed by aplant breeding technique called backcrossing, wherein essentially all ofthe morphological and physiological characteristics of a variety arerecovered in addition to the single locus transferred into the varietyvia the backcrossing technique. By essentially all of the morphologicaland physiological characteristics, it is meant that the characteristicsof a plant are recovered that are otherwise present when compared in thesame environment, other than an occasional variant trait that mightarise during backcrossing or direct introduction of a transgene.

Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the present variety. The parentalspinach plant which contributes the locus for the desired characteristicis termed the nonrecurrent or donor parent. This terminology refers tothe fact that the nonrecurrent parent is used one time in the backcrossprotocol and therefore does not recur. The parental spinach plant towhich the locus or loci from the nonrecurrent parent are transferred isknown as the recurrent parent as it is used for several rounds in thebackcrossing protocol.

In a typical backcross protocol, the original variety of interest(recurrent parent) is crossed to a second variety (nonrecurrent parent)that carries the single locus of interest to be transferred. Theresulting progeny from this cross are then crossed again to therecurrent parent and the process is repeated until a spinach plant isobtained wherein essentially all of the morphological and physiologicalcharacteristics of the recurrent parent are recovered in the convertedplant, in addition to the single transferred locus from the nonrecurrentparent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalvariety. To accomplish this, a single locus of the recurrent variety ismodified or substituted with the desired locus from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphologicalconstitution of the original variety. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross; one ofthe major purposes is to add some commercially desirable trait to theplant. The exact backcrossing protocol will depend on the characteristicor trait being altered and the genetic distance between the recurrentand nonrecurrent parents. Although backcrossing methods are simplifiedwhen the characteristic being transferred is a dominant allele, arecessive allele, or an additive allele (between recessive anddominant), may also be transferred. In this instance it may be necessaryto introduce a test of the progeny to determine if the desiredcharacteristic has been successfully transferred.

In one embodiment, progeny spinach plants of a backcross in which aplant described herein is the recurrent parent comprise (i) the desiredtrait from the non-recurrent parent and (ii) all of the physiologicaland morphological characteristics of spinach the recurrent parent asdetermined at the 5% significance level when grown in the sameenvironmental conditions.

New varieties can also be developed from more than two parents. Thetechnique, known as modified backcrossing, uses different recurrentparents during the backcrossing. Modified backcrossing may be used toreplace the original recurrent parent with a variety having certain moredesirable characteristics or multiple parents may be used to obtaindifferent desirable characteristics from each.

With the development of molecular markers associated with particulartraits, it is possible to add additional traits into an established germline, such as represented here, with the end result being substantiallythe same base germplasm with the addition of a new trait or traits.Molecular breeding, as described in Moose and Mumm, 2008 (PlantPhysiology, 147: 969-977), for example, and elsewhere, provides amechanism for integrating single or multiple traits or QTL into an eliteline. This molecular breeding-facilitated movement of a trait or traitsinto an elite line may encompass incorporation of a particular genomicfragment associated with a particular trait of interest into the eliteline by the mechanism of identification of the integrated genomicfragment with the use of flanking or associated marker assays. In theembodiment represented here, one, two, three or four genomic loci, forexample, may be integrated into an elite line via this methodology. Whenthis elite line containing the additional loci is further crossed withanother parental elite line to produce hybrid offspring, it is possibleto then incorporate at least eight separate additional loci into thehybrid. These additional loci may confer, for example, such traits as adisease resistance or a fruit quality trait. In one embodiment, eachlocus may confer a separate trait. In another embodiment, loci may needto be homozygous and exist in each parent line to confer a trait in thehybrid. In yet another embodiment, multiple loci may be combined toconfer a single robust phenotype of a desired trait.

Many single locus traits have been identified that are not regularlyselected for in the development of a new inbred but that can be improvedby backcrossing techniques. Single locus traits may or may not betransgenic; examples of these traits include, but are not limited to,herbicide resistance, resistance to bacterial, fungal, or viral disease,insect resistance, modified fatty acid or carbohydrate metabolism, andaltered nutritional quality. These comprise genes generally inheritedthrough the nucleus.

Direct selection may be applied where the single locus acts as adominant trait. For this selection process, the progeny of the initialcross are assayed for viral resistance and/or the presence of thecorresponding gene prior to the backcrossing. Selection eliminates anyplants that do not have the desired gene and resistance trait, and onlythose plants that have the trait are used in the subsequent backcross.This process is then repeated for all additional backcross generations.

Selection of spinach plants for breeding is not necessarily dependent onthe phenotype of a plant and instead can be based on geneticinvestigations. For example, one can utilize a suitable genetic markerwhich is closely genetically linked to a trait of interest. One of thesemarkers can be used to identify the presence or absence of a trait inthe offspring of a particular cross, and can be used in selection ofprogeny for continued breeding. This technique is commonly referred toas marker assisted selection. Any other type of genetic marker or otherassay which is able to identify the relative presence or absence of atrait of interest in a plant can also be useful for breeding purposes.Procedures for marker assisted selection are well known in the art. Suchmethods will be of particular utility in the case of recessive traitsand variable phenotypes, or where conventional assays may be moreexpensive, time consuming or otherwise disadvantageous. Types of geneticmarkers which could be used in accordance with the invention include,but are not necessarily limited to, Simple Sequence Length Polymorphisms(SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990),Randomly Amplified Polymorphic DNAs (RAPDs), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified FragmentLength Polymorphisms (AFLPs) (EP 534 858, specifically incorporatedherein by reference in its entirety), and Single NucleotidePolymorphisms (SNPs) (Wang et al., Science, 280:1077-1082, 1998).

F. Plants Derived By Genetic Engineering

Many useful traits that can be introduced by backcrossing, as well asdirectly into a plant, are those which are introduced by genetictransformation techniques. Genetic transformation may therefore be usedto insert a selected transgene into a plant of the invention or may,alternatively, be used for the preparation of transgenes which can beintroduced by backcrossing. Methods for the transformation of plantsthat are well known to those of skill in the art and applicable to manycrop species include, but are not limited to, electroporation,microprojectile bombardment, Agrobacterium-mediated transformation anddirect DNA uptake by protoplasts.

To effect transformation by electroporation, one may employ eitherfriable tissues, such as a suspension culture of cells or embryogeniccallus or alternatively one may transform immature embryos or otherorganized tissue directly. In this technique, one would partiallydegrade the cell walls of the chosen cells by exposing them topectin-degrading enzymes (pectolyases) or mechanically wound tissues ina controlled manner.

An efficient method for delivering transforming DNA segments to plantcells is microprojectile bombardment. In this method, particles arecoated with nucleic acids and delivered into cells by a propellingforce. Exemplary particles include those comprised of tungsten,platinum, and preferably, gold. For the bombardment, cells in suspensionare concentrated on filters or solid culture medium. Alternatively,immature embryos or other target cells may be arranged on solid culturemedium. The cells to be bombarded are positioned at an appropriatedistance below the macroprojectile stopping plate.

An illustrative embodiment of a method for delivering DNA into plantcells by acceleration is the Biolistics Particle Delivery System, whichcan be used to propel particles coated with DNA or cells through ascreen, such as a stainless steel or Nytex screen, onto a surfacecovered with target cells. The screen disperses the particles so thatthey are not delivered to the recipient cells in large aggregates.Microprojectile bombardment techniques are widely applicable, and may beused to transform virtually any plant species.

Agrobacterium-mediated transfer is another widely applicable system forintroducing gene loci into plant cells. An advantage of the technique isthat DNA can be introduced into whole plant tissues, thereby bypassingthe need for regeneration of an intact plant from a protoplast. ModernAgrobacterium transformation vectors are capable of replication in E.coli as well as Agrobacterium, allowing for convenient manipulations(Klee et al., Bio-Technology, 3(7):637-642, 1985). Moreover, recenttechnological advances in vectors for Agrobacterium-mediated genetransfer have improved the arrangement of genes and restriction sites inthe vectors to facilitate the construction of vectors capable ofexpressing various polypeptide coding genes. The vectors described haveconvenient multi-linker regions flanked by a promoter and apolyadenylation site for direct expression of inserted polypeptidecoding genes. Additionally, Agrobacterium containing both armed anddisarmed Ti genes can be used for transformation.

In those plant strains where Agrobacterium-mediated transformation isefficient, it is the method of choice because of the facile and definednature of the gene locus transfer. The use of Agrobacterium-mediatedplant integrating vectors to introduce DNA into plant cells is wellknown in the art (Fraley et al., Bio/Technology, 3:629-635, 1985; U.S.Pat. No. 5,563,055).

Transformation of plant protoplasts also can be achieved using methodsbased on calcium phosphate precipitation, polyethylene glycol treatment,electroporation, and combinations of these treatments (see, e.g.,Potrykus et al., Mol. Gen. Genet., 199:183-188, 1985; Omirulleh et al.,Plant Mol. Biol., 21(3):415-428, 1993; Fromm et al., Nature,312:791-793, 1986; Uchimiya et al., Mol. Gen. Genet., 204:204, 1986;Marcotte et al., Nature, 335:454, 1988). Transformation of plants andexpression of foreign genetic elements is exemplified in Choi et al.(Plant Cell Rep., 13: 344-348, 1994), and Ellul et al. (Theor. Appl.Genet., 107:462-469, 2003).

A number of promoters have utility for plant gene expression for anygene of interest including but not limited to selectable markers,scoreable markers, genes for pest tolerance, disease resistance,nutritional enhancements and any other gene of agronomic interest.Examples of constitutive promoters useful for plant gene expressioninclude, but are not limited to, the cauliflower mosaic virus (CaMV)P-35S promoter, which confers constitutive, high-level expression inmost plant tissues (see, e.g., Odel et al., Nature, 313:810, 1985),including in monocots (see, e.g., Dekeyser et al., Plant Cell, 2:591,1990; Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990); a tandemlyduplicated version of the CaMV 35S promoter, the enhanced 35S promoter(P-e35S); 1 the nopaline synthase promoter (An et al., Plant Physiol.,88:547, 1988); the octopine synthase promoter (Fromm et al., Plant Cell,1:977, 1989); and the figwort mosaic virus (P-FMV) promoter as describedin U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter(P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem;the cauliflower mosaic virus 19S promoter; a sugarcane bacilliform viruspromoter; a commelina yellow mottle virus promoter; and other plant DNAvirus promoters known to express in plant cells.

A variety of plant gene promoters that are regulated in response toenvironmental, hormonal, chemical, and/or developmental signals can alsobe used for expression of an operably linked gene in plant cells,including promoters regulated by (1) heat (Callis et al., PlantPhysiol., 88:965, 1988), (2) light (e.g., pea rbcS-3A promoter,Kuhlemeier et al., Plant Cell, 1:471, 1989; maize rbcS promoter,Schaffner and Sheen, Plant Cell, 3:997, 1991; or chlorophyll a/b-bindingprotein promoter, Simpson et al., EMBO J., 4:2723, 1985), (3) hormones,such as abscisic acid (Marcotte et al., Plant Cell, 1:969, 1989), (4)wounding (e.g., wunl, Siebertz et al., Plant Cell, 1:961, 1989); or (5)chemicals such as methyl jasmonate, salicylic acid, or Safener. It mayalso be advantageous to employ organ-specific promoters (e.g., Roshal etal., EMBO J., 6:1155, 1987; Schernthaner et al., EMBO J., 7:1249, 1988;Bustos et al., Plant Cell, 1:839, 1989).

Exemplary nucleic acids which may be introduced to plants of thisinvention include, for example, DNA sequences or genes from anotherspecies, or even genes or sequences which originate with or are presentin the same species, but are incorporated into recipient cells bygenetic engineering methods rather than classical reproduction orbreeding techniques. However, the term “exogenous” is also intended torefer to genes that are not normally present in the cell beingtransformed, or perhaps simply not present in the form, structure, etc.,as found in the transforming DNA segment or gene, or genes which arenormally present and that one desires to express in a manner thatdiffers from the natural expression pattern, e.g., to over-express.Thus, the term “exogenous” gene or DNA is intended to refer to any geneor DNA segment that is introduced into a recipient cell, regardless ofwhether a similar gene may already be present in such a cell. The typeof DNA included in the exogenous DNA can include DNA which is alreadypresent in the plant cell, DNA from another plant, DNA from a differentorganism, or a DNA generated externally, such as a DNA sequencecontaining an antisense message of a gene, or a DNA sequence encoding asynthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and couldpotentially be introduced into a spinach plant according to theinvention. Non-limiting examples of particular genes and correspondingphenotypes one may choose to introduce into a spinach plant include oneor more genes for insect tolerance, such as a Bacillus thuringiensis(B.t.) gene, pest tolerance such as genes for fungal disease control,herbicide tolerance such as genes conferring glyphosate tolerance, andgenes for quality improvements such as yield, nutritional enhancements,environmental or stress tolerances, or any desirable changes in plantphysiology, growth, development, morphology or plant product(s). Forexample, structural genes would include any gene that confers insecttolerance including but not limited to a Bacillus insect control proteingene as described in WO 99/31248, herein incorporated by reference inits entirety, U.S. Pat. No. 5,689,052, herein incorporated by referencein its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, hereinincorporated by reference in their entirety. In another embodiment, thestructural gene can confer tolerance to the herbicide glyphosate asconferred by genes including, but not limited to Agrobacterium strainCP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat.No. 5,633,435, herein incorporated by reference in its entirety, orglyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No.5,463,175, herein incorporated by reference in its entirety.

Alternatively, the DNA coding sequences can affect these phenotypes byencoding a non-translatable RNA molecule that causes the targetedinhibition of expression of an endogenous gene, for example viaantisense- or cosuppression-mediated mechanisms (see, for example, Birdet al., Biotech. Gen. Engin. Rev., 9:207, 1991). The RNA could also be acatalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desiredendogenous mRNA product (see for example, Gibson and Shillito, Mol.Biotech., 7:125, 1997). Thus, any gene which produces a protein or mRNAwhich expresses a phenotype or morphology change of interest is usefulfor the practice of the present invention.

G. Definitions

In the description and tables herein, a number of terms are used. Inorder to provide a clear and consistent understanding of thespecification and claims, the following definitions are provided:

Allele: Any of one or more alternative forms of a gene locus, all ofwhich alleles relate to one trait or characteristic. In a diploid cellor organism, the two alleles of a given gene occupy corresponding locion a pair of homologous chromosomes.

Backcrossing: A process in which a breeder repeatedly crosses hybridprogeny, for example a first generation hybrid (F₁), back to one of theparents of the hybrid progeny. Backcrossing can be used to introduce oneor more single locus conversions from one genetic background intoanother.

Crossing: The mating of two parent plants.

Cross-pollination: Fertilization by the union of two gametes fromdifferent plants.

Diploid: A cell or organism having two sets of chromosomes.

Emasculate: The removal of plant male sex organs or the inactivation ofthe organs with a cytoplasmic or nuclear genetic factor or a chemicalagent conferring male sterility.

Enzymes: Molecules which can act as catalysts in biological reactions.

F₁ Hybrid: The first generation progeny of the cross of two nonisogenicplants.

Genotype: The genetic constitution of a cell or organism.

Haploid: A cell or organism having one set of the two sets ofchromosomes in a diploid.

Linkage: A phenomenon wherein alleles on the same chromosome tend tosegregate together more often than expected by chance if theirtransmission was independent.

Marker: A readily detectable phenotype, preferably inherited incodominant fashion (both alleles at a locus in a diploid heterozygoteare readily detectable), with no environmental variance component, i.e.,heritability of 1.

Phenotype: The detectable characteristics of a cell or organism, whichcharacteristics are the manifestation of gene expression.

Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer togenetic loci that control to some degree numerically representabletraits that are usually continuously distributed.

Resistance: As used herein, the terms “resistance” and “tolerance” areused interchangeably to describe plants that show no symptoms to aspecified biotic pest, pathogen, abiotic influence or environmentalcondition. These terms are also used to describe plants showing somesymptoms but that are still able to produce marketable product with anacceptable yield. Some plants that are referred to as resistant ortolerant are only so in the sense that they may still produce a crop,even though the plants are stunted and the yield is reduced.

Regeneration: The development of a plant from tissue culture.

Royal Horticultural Society (RHS) Color Chart Value: The RHS color chartis a standardized reference which allows accurate identification of anycolor. A color's designation on the chart describes its hue, brightnessand saturation. A color is precisely named by the RHS color chart byidentifying the group name, sheet number and letter, e.g., Yellow-OrangeGroup 19A or Red Group 41B.

Self-pollination: The transfer of pollen from the anther to the stigmaof the same plant.

Single Locus Converted (Conversion) Plant: Plants which are developed bya plant breeding technique called backcrossing, wherein essentially allof the morphological and physiological characteristics of a spinachvariety are recovered in addition to the characteristics of the singlelocus transferred into the variety via the backcrossing technique and/orby genetic transformation.

Substantially Equivalent: A characteristic that, when compared, does notshow a statistically significant difference (e.g., p=0.05) from themean.

Tissue Culture: A composition comprising isolated cells of the same or adifferent type or a collection of such cells organized into parts of aplant.

Transgene: A genetic locus comprising a sequence which has beenintroduced into the genome of a spinach plant by transformation.

H. Deposit Information

A deposit of spinach hybrid SV2146VB and inbred parent lines SSB66-1129Fand ESA66-1128M, disclosed above and recited in the claims, has beenmade with the American Type Culture Collection (ATCC), 10801 UniversityBlvd., Manassas, Va. 20110-2209. The date of deposits were Jul. 17,2013, Aug. 2, 2011, and Jul. 17, 2013, respectively. The accessionnumber for those deposited seeds of spinach hybrid SV2146VB and inbredparent lines SSB66-1129F and ESA66-1128M, are ATCC Accession No.PTA-120468, ATCC Accession No. PTA-12019, and ATCC Accession No.PTA-120469, respectively. Upon issuance of a patent, all restrictionsupon the deposits will be removed, and the deposits are intended to meetall of the requirements of 37 C.F.R. §1.801-1.809. The deposits will bemaintained in the depository for a period of 30 years, or 5 years afterthe last request, or for the effective life of the patent, whichever islonger, and will be replaced if necessary during that period.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity andunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the invention, as limited only bythe scope of the appended claims.

All references cited herein are hereby expressly incorporated herein byreference.

What is claimed is:
 1. A spinach plant comprising at least a first setof the chromosomes of spinach line ESA66-1128M, a sample of seed of saidline having been deposited under ATCC Accession Number PTA-120469.
 2. Aseed comprising at least a first set of the chromosomes of spinach lineESA66-1128M, a sample of seed of said line having been deposited underATCC Accession Number PTA-120469.
 3. The plant of claim 1, which is aninbred.
 4. The plant of claim 1, which is a hybrid.
 5. The seed of claim2, which is an inbred.
 6. The seed of claim 2, which is a hybrid.
 7. Theplant of claim 4, wherein the hybrid plant is spinach hybrid SV2146VB, asample of seed of said hybrid SV2146VB having been deposited under ATCCAccession Number PTA-120468.
 8. The seed of claim 6, wherein the seed isa seed of spinach hybrid SV2146VB, a sample of seed of said hybridSV2146VB having been deposited under ATCC Accession Number PTA-120468.9. A plant part of the plant of claim
 1. 10. The plant part of claim 9,wherein the plant part is a fruit, an ovule, pollen, a leaf, or a cell.11. A spinach plant having all the physiological and morphologicalcharacteristics of the spinach plant of claim
 7. 12. A tissue culture ofregenerable cells of the plant of claim
 1. 13. The tissue cultureaccording to claim 12, comprising cells or protoplasts from a plant partselected from the group consisting of embryos, meristems, cotyledons,pollen, leaves, anthers, roots, root tips, pistil, flower, seed andstalks.
 14. A spinach plant regenerated from the tissue culture of claim12, wherein said plant comprises all of the morphological andphysiological characteristics of the spinach plant comprising at least afirst set of the chromosomes of spinach line ESA66-1128M, a sample ofseed of said line having been deposited under ATCC Accession NumberPTA-120469.
 15. A method of vegetatively propagating the spinach plantof claim 1 comprising the steps of: (a) collecting tissue capable ofbeing propagated from the plant according to claim 1; (b) cultivatingsaid tissue to obtain proliferated shoots; and (c) rooting saidproliferated shoots to obtain a rooted plantlet.
 16. The method of claim15, further comprising growing at least a first spinach plant from saidrooted plantlet.
 17. A method of introducing one desired trait intospinach line ESA66-1128M comprising: (a) utilizing as a recurrent parenta plant of spinach line ESA66-1128M, by crossing a plant of lineESA66-1128M with a second donor spinach plant that comprises the desiredtrait to produce F1 progeny, a sample of seed of said line having beendeposited under ATCC Accession Number PTA-120469; (b) selecting an F1progeny that comprises the desired trait; (c) backcrossing the selectedF1 progeny with a plant of line ESA66-1128M to produce backcrossprogeny; (d) selecting backcross progeny comprising the desired trait;and (e) repeating steps (c) and (d) three or more times to produce aselected fourth or higher backcross progeny that comprise the desiredtrait, and otherwise comprises all of the morphological andphysiological characteristics of spinach line ESA66-1128M.
 18. A spinachplant produced by the method of claim
 17. 19. A method of producing aspinach plant comprising an added trait, the method comprisingintroducing a transgene conferring the trait into a plant of hybridSV2146VB, or line ESA66-1128M, a sample of seed of said hybrid and linehaving been deposited under ATCC Accession Number PTA-120468, and ATCCAccession Number PTA-120469, respectively.
 20. A spinach plant producedby the method of claim
 19. 21. The plant of claim 1, further comprisinga transgene.
 22. The plant of claim 21, wherein the transgene confers atrait selected from the group consisting of male sterility, herbicidetolerance, insect resistance, pest resistance, disease resistance,modified fatty acid metabolism, environmental stress tolerance, modifiedcarbohydrate metabolism and modified protein metabolism.
 23. The plantof claim 1, further comprising a single locus conversion.
 24. The plantof claim 23, wherein the single locus conversion confers a traitselected from the group consisting of male sterility, herbicidetolerance, insect resistance, pest resistance, disease resistance,modified fatty acid metabolism, environmental stress tolerance, modifiedcarbohydrate metabolism and modified protein metabolism.
 25. A methodfor producing a seed of a spinach plant derived from at least one ofhybrid SV2146VB, or line ESA66-1128M comprising the steps of: (a)crossing a spinach plant of hybrid SV2146VB, or line ESA66-1128M withitself or a second spinach plant; a sample of seed of said hybrid andline having been deposited under ATCC Accession Number PTA-120468, andATCC Accession Number PTA-120469, respectively; and (b) allowing a seedof the hybrid SV2146VB-derived, or line ESA66-1128M-derived spinachplant to form.
 26. The method of claim 25, further comprising the stepsof: (c) selfing a plant grown from said hybrid SV2146VB-derived, orESA66-1128M-derived spinach seed to yield an additional hybridSV2146VB-derived, or line ESA66-1128M-derived spinach seed; (d) growingsaid additional hybrid SV2146VB-derived, or line ESA66-1128M-derivedspinach seed of step (c) to yield an additional hybrid SV2146VB-derived,or line ESA66-1128M-derived spinach plant; and (e) repeating thecrossing and growing steps of (c) and (d) to generate at least a firstfurther hybrid SV2146VB-derived, or line ESA66-1128M-derived spinachplant.
 27. The method of claim 25, wherein the second spinach plant isof an inbred spinach line.
 28. The method of claim 25, comprisingcrossing line ESA66-1128M with line SSB 66-1129F, a sample of seed ofsaid lines having been deposited under ATCC Accession Number PTA-120469,and ATCC Accession Number PTA-12019, respectively.
 29. The method ofclaim 26, further comprising: (f) crossing the further hybridSV2146VB-derived, or ESA66-1128M-derived spinach plant with a secondspinach plant to produce a seed of a hybrid progeny plant.
 30. A plantpart of the plant of claim
 7. 31. The plant part of claim 30, whereinthe plant part is a fruit, an ovule, pollen, a leaf, or a cell.
 32. Amethod of producing a spinach seed comprising crossing the plant ofclaim 1 with itself or a second spinach plant and allowing seed to form.33. A method of producing a spinach fruit comprising: (a) obtaining aplant according to claim 1, wherein the plant has been cultivated tomaturity; and (b) collecting a spinach fruit from the plant.
 34. Amethod of producing a plant of spinach hybrid SV2146VB comprising asingle locus conversion, the method comprising crossing a plant of lineSSB 66-1129F with a plant of line ESA66-1128M, wherein one of said linescomprises a single locus conversion, a sample of seed of said lines SSB66-1129F and ESA66-1128M having been deposited under ATCC Accession No.PTA-12019 and ATCC Accession Number PTA-120469, respectively.